Re: Intracellular/extracellular cytokine detection using GAM-PE secondary AB

From: Art Roberts (robertar@UMDNJ.EDU)
Date: Tue Mar 16 1999 - 11:52:09 EST


Greg,

I can help you with the last question.  Your human AB serum probably has
complement activity which will definitely lyse any antibody labelled
cells.  Heat de-activation will usually take care of this.  But I would
recommend using serum from the host animal in which the antibody was
raised-mouse or goat.  It should more effectively block any non-specific
binding.

Art  

On Thu, 11 Mar 1999, Greg Neely wrote:

> 
>
> 5. I just tried to stain my cells for surface markers in PBS, 0.1%
> sodium azide, but 1% human AB serum instead of fetal calf serum, hoping
> this might better block non-specific interactions.  Instead my cell
> count was absurdly low, and there was tons of dead cells.  Has anyone
> seen this?  Is this why one does not traditionally wash/stain cells in
> human serum?
> 
> 
> 

Art Roberts
Dept. of Medicine
Robert Wood Johnson Medical School

email:  robertar@umdnj.edu
phone:  732-235-7790
Fax:    732-235-7792



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